用巢式PCR方法制备基因芯片的探针  

Preparation of Microarray Probes Using Nested PCR Approaches

作　　者：黄海[1] 马文丽[1] 王洪敏[1] 肖维威[1] 王艳[1] 郑文岭[2] 

机构地区：[1]第一军医大学分子生物学研究所,中国广东广州510515 [2]广州军区总医院医学实验科,中国广东广州510010

出　　处：《生命科学研究》2003年第3期271-274,共4页Life Science Research

基　　金：广州市重点科技攻关项目(990448022)

摘　　要：制备出高纯度的探针,用于诊断基因芯片的打印.采用巢式PCR技术,M13作为外侧引物并自行设计内侧引物,扩增克隆在T载体上的基因片段.可制备出成分单一,上下游仅含19bp和20bp的短载体序列的探针,能使打印出的芯片得到较好的杂交效果.该法充分利用巢式PCR的优点,对制备探针的方法进行改进,且简便快速,能得到更高质量的探针,满足打印芯片的要求.Probes with high purity were amplified by nested PCR in DNA microarray preparations. Nested PCR was applied to amplify the clones in PMD 18T vector with M13 forward and reverse primers as external primers, while a pair of inner primers were designed. Highquality probes could be isolated containing short Tvector sequence. The DNA microarray, which printed with these probes, showed superior hybridization results. The advantages of nested PCR improved the efficiency of preparing probes to make diagnostic microarrays. 

关 键 词：巢式PCR 基因芯片 探针 T载体 

分 类 号：Q78[生物学—分子生物学] Q503